The Library
An optimized SEC-SAXS system enabling high X-ray dose for rapid SAXS assessment with correlated UV measurements for biomolecular structure analysis
Tools
Ryan, Timothy M., Trewhella, Jill, Murphy, James M., Keown, Jeremy R., Casey, Lachlan, Pearce, F. Grant, Goldstone, David C., Chen, Kelan, Luo, Zhenyao, Kobe, Bostjan, McDevitt, Christopher A., Watkin, Serena A., Hawley, Adrian M., Mudie, Stephen T., Samardzic Boban, Vesna and Kirby, Nigel (2018) An optimized SEC-SAXS system enabling high X-ray dose for rapid SAXS assessment with correlated UV measurements for biomolecular structure analysis. Journal of Applied Crystallography, 51 (1). pp. 97-111. doi:10.1107/S1600576717017101 ISSN 0021-8898.
Research output not available from this repository.
Request-a-Copy directly from author or use local Library Get it For Me service.
Official URL: http://doi.org/10.1107/S1600576717017101
Abstract
A new optimized size exclusion chromatography small-angle X-ray scattering (SEC-SAXS) system for biomolecular SAXS at the Australian Synchrotron SAXS/WAXS beamline has been developed. The compact configuration reduces sample dilution to maximize sensitivity. Coflow sample presentation allows an 11-fold increase in flux on sample without capillary fouling, improving throughput and data quality, which are now primarily limited by the full flux available on the beamline. Multi-wavelength fibre optic UV analysis in close proximity to the X-ray beam allows for accurate concentration determination for samples with known UV extinction coefficients and thus estimation of the molecular weight of the scattering particle from the forward X-ray scattering intensity. Fast-flow low-volume SEC columns provide sample throughput competitive with batch concentration series measurements, albeit with a concomitant reduction of potential resolution relative to lower flow rates and larger SEC columns. The performance of the system is demonstrated using a set of model proteins, and its utility to solve various challenges is illustrated with a diverse suite of protein samples. These developments increase the quality and rigor of SEC-SAXS analysis and open new avenues for biomolecular solution SEC-SAXS studies that have been challenged by low sample yields, temporal instability, radiation sensitivity and complex mixtures.
Item Type: | Journal Article | ||||
---|---|---|---|---|---|
Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||
Journal or Publication Title: | Journal of Applied Crystallography | ||||
Publisher: | Wiley-Blackwell Publishing Ltd. | ||||
ISSN: | 0021-8898 | ||||
Official Date: | 1 February 2018 | ||||
Dates: |
|
||||
Volume: | 51 | ||||
Number: | 1 | ||||
Page Range: | pp. 97-111 | ||||
DOI: | 10.1107/S1600576717017101 | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published | ||||
Access rights to Published version: | Restricted or Subscription Access |
Request changes or add full text files to a record
Repository staff actions (login required)
View Item |